1.Quality standard study on Pteris multifida.
Jian-qun LIU ; Jian LIU ; Wei JIANG ; Guo-yue ZHONG
China Journal of Chinese Materia Medica 2015;40(10):1855-1859
The quality control method and standard were established to control the quality of Pteris multifida in this paper. The tests of water content, total ash, acid-unsoluble ash and ethanol-soluble extractives of P. multifida were carried out according to the methods recorded in appendix of Chinese Pharmacopeia (2010 edition, volume 1) . The TLC method was established by using rhoifolin as references, and a mixture of CHCl3 -MeOH-HAc (6: 1: 1) as the developing solvent system on GF254 thin layer plate. The contents of rhoifolin was determined by HPLC on a Diamonsil C18 (4.6 mm x 250 mm, 5 μm) column, using acetonitrile-water (containing 0.15% formic acid) (16: 84) as mobile phase at a flow rate of 1.0 mL x min(-1). The column temperature was 30 degrees C and the detection wave-length was 350 nm. As a result, pterosin C 3-O-β-D-glucosidede and the other constituents were well separated on TLC detected under the UV light at 254 nm . The methodology validation for the assay of rhoifolin presented that it was in good linear correlation in the ranges of 0.025 5-5.1 μg with the regression equations of Y = 1 092.4X + 9.503 5 (r = 0.999 8), and the average recoveries were 100.3% (RSD 1.3%). The content range of rhoifolin from 16 different batches of Pteris multifida was 0.08-5.06 mg x g(-1). The water content, total ash, acid-unsoluble ash and ethanol-soluble extractives of 16 samples varied in the ranges of 7.35% - 12.96%, 6.90% - 16.33%, 2.07% -11.38% and 13.29% -23.87%, respectively. The suggesting limes in the quality standard for water content, total ash, acid-unsoluble ash, ethanol-soluble extractives and rhoifolin content were ≤ 12% , ≤ 15% , ≤ 8.5% , ≥ 14% and ≥ 0.040%, respectively. The result proved that the established quality of control method was specific and accurate, which can be used for the quality control of P. multifida.
China
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Chromatography, High Pressure Liquid
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Chromatography, Thin Layer
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Drugs, Chinese Herbal
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chemistry
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isolation & purification
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standards
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Pteris
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chemistry
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Quality Control
2.Biological features and ultrastructure of rabbit mesenchymal stem cells in vitro
Jie LIU ; Jian WANG ; Yue ZHOU
Medical Journal of Chinese People's Liberation Army 2001;0(10):-
Objective To isolate and culture the rabbit mesenchymal stem cells,and explore the biological features in vitro.Methods Two milliliter of bone marrow was extracted from the tibia of juvenile rabbit under anesthesia and sterile conditions,and mesenchymal stem cells were obtained by density gradient centrifugation and inoculated to 100ml culture flask.The morphological changes,adherence rate and ultrastructure of the stem cells were observed.The surface markers and growth cycle of the mesenchymal stem cells were detected by flow cytometry,and biological features were observed by inducing the cells differentiate into osteoblasts.Results The adherence of mesenchymal stem cells started 2 hours after culturing,basically finished 20 hours later,and more than 90% of the cells were fused at the 15th day.Observed with transmission electron microscopy,the cultured cells showed typical ultrastructure of stem cells: numbers of microvillus mainly distributed on one side of the cells,irregular nuclei with indentation,and lots of cell organs.Analysis for the cell surface markers indicated positive expression of CD44 and CD90 and negative expression of CD45 in mesenchymal stem cells.Cell growth cycle assay showed that most mesenchymal stem cells(about 97%) stayed in stage G1 and G2.Three weeks after osteoblastic induction,the mesenchymal stem cells gradually produced small calcium salt crystals,the cells were positive in alkaline phosphatase staining,and Von Kossa staining showed many calcium salts in the cell clone as black reaction deposits.Conclusions The rabbit bone marrow mesenchymal stem cells isolated by density gradient centrifugation are of high purity and stable growth rate,and the cells' characters can be still maintained after serial subcultivation.Since easy to operate on isolation and proliferation,the mesenchymal stem cells could be used as the tissue engineering seed cells in repairing of intervertebral disc degeneration.
3.Construction of lentiviral vector miR30-CD133 targeting CD133 gene and its inhibition of liver cancer cell growth
The Journal of Practical Medicine 2016;32(10):1571-1574
Objective To construct lentiviral vector miR30-CD133 targeting CD133 gene and investigate its effects on SMMC7721 cells. Methods Thespecific sequence of DNA which containing both sense and antisense Oligo DNA of the targeting sequence were designed, synthesized and cloned into the pPRIME vector. pPRIME-miR30-CD133, psPAX2 and pMD2G were co-transfected on 293T cells to get the lentivirus the transfection efficiency was investigated by confocal laser scanning microscope. The expression of miR30-CD133 on CD133 were detected by qRT-PCR and Western blott. The proliferation and apoptosis effect of miR30-CD133 was respectively evaluated by CCK-8 assay and AnnexinV/PI. Results Functional PFU titers of PRIME-miR30-CD133 were 6.58 × 109 PFU/mL. The expression of CD133 mRNA in sh CD133 group (0.18 ± 0.04) was less than Blank group and shNC group (P<0.05). The expression of CD133 protein in sh CD133 group was less than Blank group (10%) and shNC group (8%) (P < 0.05). Cells proliferation activity were significantly inhibited in shCD133 group compared with control group. Apoptosis rate were significantly increased in shCD133 group (41.3%) compared with Blank group (25.3%)and shNC group (24.3%). Conclusion The lentivirus miR30 vector targeting CD133 gene was successfully constructed, which will be a basis to the further CD133 gene studies.
4.Evaluation of oral gastrointestinal ultrasound contrast agents on gastric motility in patients with anxiety disorder
Wenyan LI ; Peng GU ; Wensheng YUE ; Jian LIU ; Shudong LIU
Chongqing Medicine 2016;45(20):2798-2800
Objective To apply the oral gastrointestinal ultrasound contrast agents for evaluating the gastric motility abnor‐mality in the patients with anxiety disorder .Methods Twenty patients with anxiety disorder complicating upper digestive tract symptoms without organic pathological changes were enrolled as the anxiety disorder group .Twenty healthy volunteers were en‐rolled as the control group in this study .The two groups orally took gastrointestinal ultrasound contrast agents .The antral contrac‐tion frequency ,antral contraction amplitude amd MI were measured at each time point .GER was calculated .The the gastric motility parameters in the patients with anxiety disorder were evaluated .Results The antral contraction frequency and MI at initial 2 min had no statistical difference between the anxiety group and the control group .The antral contraction amplitude ,antral contraction frequency amd MI at each time point during 5 -10 min after contrast in the anxiety disorder group were significantly decreased compared with the control group ,and the differences were statistically significant (P< 0 .05) .After 20 min ,GER in the control group was significantly higher than that in the anxiety group ,the difference was statistically significant (P<0 .05) .Conclusion O‐ral gastrointestinal ultrasound contrast agents is a economic ,strongly operable ,non‐invasive and highly repeatable method for evalu‐ating the gastric motility .
5.Effect of ginsenoside total saponinon on regulation of P450 of livers of rats after γ-ray irradiation.
Jian-feng YI ; Bo WU ; Cang-long LIU ; Yue GAO
China Journal of Chinese Materia Medica 2015;40(20):4037-4043
Effect of ginsenoside total saponin (GTS) on the regulation of P450 of livers of rats after γ-ray irradiation was studied. Rats were irradiated by the ⁶⁰Coγ-ray for one-time dose of 5.5 Gy, dose rate of 117.1-119.2 cGy. The cocktail probe, qPCR and Western blot were used to detect expression of enzymatic activites, mRNA and protein of rats. Contrasted with blank group, expression of CYP1A2, 2B1, 2E1, 3A4 of irradiation group showed a up-regulated (P < 0.05). Contrasted with irradiation group, exprression of CYP1A2, 2B1, 2E1, 3A4 of GTS group showed a downward trend. GTS had negative agonistic action against expression of P450 of rats by irradiatied.
Animals
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Cytochrome P-450 Enzyme System
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genetics
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metabolism
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Drugs, Chinese Herbal
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pharmacology
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Gamma Rays
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Ginsenosides
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pharmacology
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Liver
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drug effects
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enzymology
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radiation effects
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Male
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Microsomes, Liver
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drug effects
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enzymology
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Panax
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chemistry
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Rats
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Rats, Wistar
6.Research progress in the structure and function of dengue virus non-structural 1 protein.
Yue CHEN ; Rui-wen REN ; Jian-wei LIU
Chinese Journal of Virology 2014;30(6):683-688
Dengue virus (DENV) is a re-emerging disease transmitted by the Aedes mosquitoes and has become a major public health problem in southern China. Currently, no antiviral drug or effective vaccine exist to control this disease. The chimeric DENV structural protein vaccine cannot elicit balanced levels of protective immunity to each of the four viral serotypes; therefore, non-structural protein components may be required to construct an effective DENV vaccine. The Dengue virus non-structural 1 (DENV NS1) protein plays a critical role in viral pathogenesis and protective immunity. Therefore, immunity to Dengue 1-4 NS1 subtypes may be crucial for the prevention of severe disease. This review attempts to provide an overview about the structure and function of DENV NS1.
Animals
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Dengue
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immunology
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prevention & control
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virology
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Dengue Vaccines
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chemistry
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genetics
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immunology
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Dengue Virus
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chemistry
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genetics
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immunology
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Humans
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Viral Nonstructural Proteins
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chemistry
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genetics
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immunology
8.3-D finite element study on maxillary body protraction using implant anchorage
Yang QU ; Jian LIU ; Siwei YANG ; Yue HUANG
Journal of Practical Stomatology 2016;32(1):58-62
Objective:To explore the stress change rule of craniofacial bone suture and the interface of bone-implant against differ-ent strength and direction of protraction on the implant anchorage in alveolar bone.Methods:The original DICOMdata of 2-D image of craniofacial complex were obtained by high resolution CT scanning.3-D finite element models of craniofacial complex were devel-oped according to the DICOMdata.Forces of 1 -1 0 N inclined at 0 -60°to Frankfort horizontal plane in the anterior and inferior di-rections were respectively applied on the implant anchorage in the alveolar bone at 32 23 .Data of principle stress and Von Mises Stress of each mode of each simulaton was caculated.Results:The change rule of the effectiveness of different force value of protrac-tion in the same direction was the same in different stress zone;that of the same force value of the protraction in differente direction differed in different stress zone.When the protraction angle was less than 30°,the maxillary complex will spin up.In the 30°,the maxillary complex showed the forward growth.Between 40°-50°,the growth direction was the same with the protraction direction. When the protraction angle was more than 50°,the maxillary complex showed down spin.Conclusion:Protraction force of 1 -1 0 N at 30°-50°to Frankfort horizontal plane on implant anchorage in the alveolar bone at 32 23 can induce maxillary complex grow for-ward.
9.Comparison of clinical features and prognosis of patients with severe traumatic brain injury among different age groups
Xiaonian ZHANG ; Jian LIU ; Yue ZHANG ; Hao ZHANG
Chinese Journal of Geriatrics 2014;33(7):726-728
Objective To investigate the influence of age on clinical features and prognosis of severe traumatic brain injury.Methods A total of 135 patients with severe traumatic brain injury were divided into four groups according to age:the juvenile group (<18 years,15 cases),the young adult group (18 44 years,77 cases),the middle aged group (45 59 years,37 cases) and the elderly group (>60 years,6 cases).Neurological functions were assessed by the Disability Rating Scale (DRS),the Mini-Mental Status Examination (MMSE),the Fugl-Meyer motor scale (FM) and the Modified Barthel Index (MBI).All the patients were followed up with DRS evaluation 1 4 years after discharge from the hospital.Results MMSE scores decreased with age,with statistically significant differences between the elderly group and the juvenile group,the young adult group and the middle aged group,respectively [(11.0±5.2) vs (21.5±8.1),(21.4±8.0) and (19.1±8.1),respectively; t=2.663、2.825、2.561,P=0.015,0.006,0.022,respectively].Similarly,when compared with the other groups,the elderly group also showed statistically significant differences in follow-up DRS scores [(12.8±6.1) vs.(4.3±3.6),(6.7±5.0) and (7.8±6.9),respectively; t=-2.382、-2.587、-2.385,P =0.040,0.013 and 0.038,respectively]; and the DRS score differentials [(2.3±4.6) vs.(6.2±4.3),(6.7±3.1) and (4.6±3.1); t=2.366、2.242、2.626,P =0.004,0.013 and 0.009,respectively].Conclusions Age may be one of the factors that affect cognitive functions and prognosis associated with traumatic brain injury,and the prognosis for elderly patients is generally unfavorable.
10.Culture and biological characteristics of neural stem cells from caveolin-1 knockout embryonic mice
Baiyan LIU ; Yue YU ; Jian YI ; Xuemei CHEN ; Guangxian CAI
Chinese Journal of Tissue Engineering Research 2014;(23):3739-3744
BACKGROUND:Caveolin-1 is expressed in mammalian brain and involved in the normal development of the brain, which can affect the proliferation of neural stem cells in the brain. OBJECTIVE:To acquire neural stem cells from caveolin-1 knockout embryonic mice in vitro and study their biological characteristics. METHODS:The whole brain was separated from C57BL/6 mice and caveolin-1 knockout C57BL/6 mice respectively at encyesis 14-16 days. Single cellsuspension was obtained by enzyme digestion, and cultured in the conditioned medium of neural stem cells. Fol owing 7 days of primary culture, the cells were induced in Dulbecco’s modified Eagle’s medium/Ham’s nutrient mixture F-12 containing 10%fetal bovine serum for 7 days. RESULTS AND CONCLUSION:The major cells of the cellsuspensions from the fetal mouse brain were dead at 1 day after culture, and some single cells floated in the medium and their transmittance were better, and then they gradual y formed multicellular bal s after 3 days. A smal amount of cells were adhered at the bottom of culture plate after passage, and a great amount of cellbal s appeared after 7 days. The proliferation rate of neural stem cells from caveolin-1 knockout mice was higher than that from normal mice. The cellbal s were nestin-positive and their differentiated cells was positive for neurofilament 200, glial fibril ary acidic protein or O4, respectively. Al of the cells from normal mouse brain were positive for caveolin-1, but the cells from caveolin-1 knockout mice were negative for caveolin-1 by immunocytochemistry. Moreover, the speed of cellbal formation and the number of cellbal s in neural stem cells from caveolin-1 knockout mice were better than those from normal mice. Caveolin-1 negative neural stem cells were cultured successful y from caveolin-1 knockout mouse brain, and the results show that caveolin-1 can promote the proliferation of neural stem cells and inhibit their differentiation in vitro.